Invitrogen

Paxillin Monoclonal Antibody (5H11)

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

12 Published Figures

22 References

View all (53) Paxillin antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

Paxillin Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Paxillin was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Paxillin (5H11) Mouse Monoclonal Antibody (Product # AHO0492) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300).Panel d is a merged image showing punctuated cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Paxillin Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of Paxillin in formalin-fixed, paraffin-embedded human colon carcinoma using a Paxillin monoclonal antibody (Product # AHO0492). Detection was performed with a peroxidase-conjugate and AEC chromogen. Note nuclear staining of tumor cells.

Paxillin Antibody in Western Blot (WB) — Knockout of Paxillin was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1071991_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Paxillin was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) andHeLa Paxillin KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-Paxillin Monoclonal Antibody (5H11) (Product # AHO0492, 1 µg/mL dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Paxillin.

Paxillin Antibody in Immunoprecipitation (IP) — Immunoprecipitation of Paxillin in native human A431 cells using a Paxillin monoclonal antibody (Product # AHO0492).

Paxillin Antibody in Immunocytochemistry, Immunohistochemistry (ICC/IF, IHC) — Fig. 5 Plexin-B2 influences cell-matrix adhesion and controls contractility in hESC colonies. a , b TIRF microscopy images and quantification show altered number, size, and shape of focal adhesions (FAs) in PLXNB2 mutant hESC colonies ( a ) and hNPCs ( b ) compared to wild-type (WT) cells. Cells were stained for FA marker paxillin and F-actin (phalloidin). Enlarged images of boxed areas are shown below in inverted black and white, with dashed lines outlining individual cells. Box plots depict 25-75% quantiles, minimal and maximal values (whiskers), median (midline), and mean (cross signs). For hESCs: FA number, n = 14-15 cells per group. FA area, n = 2271 FAs for WT, n = 1042 for PB2 KO, and n = 978 for PB2 OE. FA circularity, n = 1042 FAs for WT, n = 1042 for PB2 KO, and n = 978 for PB2 OE. * P = 0.0397, *** P < 0.0001. For hNPCs: FA number, n = 20 cells per group. FA area and circularity, n = 1829 FAs for WT, n = 2385 for PB2 KO, and n = 4528 for PB2 OE. Kruskal-Wallis test followed by Dunn's multiple comparisons test. ** P = 0.0084, *** P < 0.0001. c Molecular dynamics mathematical simulation. Left, components of simulation based on a bead-spring model. The tip of the simulated AFM probe is depicted as a green sphere, with elastic constants K x and K y in the x and y directions, respectively. Right, graphs of force as a function of distance traveled by the simulated AFM probe in two different scenarios. Left, when varying the actin-membrane attraction energy that simulates

Paxillin Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Figure 6 FAK-paxillin interaction conditions FAK localization to FA. ( A ) Cell extracts from A375 melanoma cells transiently co-transfected with siFAK and wild-type FAK-GFP or FAK-I936/I998-GFP were analyzed by immunoprecipitation (IP) using anti-FAK Ab and blotted for paxillin and FAK. The expression level of proteins in the corresponding cell lysate is shown. ( B ) Representative Western blot showing A375 melanoma cells transfected as described above and blotted for P-Y397 FAK, FAK total, eGFP and actin. ( C ) Cells transiently co-transfected with siFAK and wild-type FAK-GFP or FAK-I936/I998-GFP were fixed and labelled for paxillin (purple), actin (red) and P-Y397 FAK (cyan). Scale bar: 10 um. ( D ) Histograms represent the mean normalized to control +- SD of FAK localization in melanoma cells. Signals were measured by reporting the P-FAK signal in FA over that in the cytoplasm from 3 independent experiments ***, p < 0.001; unpaired t -test compared to control condition.

Paxillin Antibody in Immunohistochemistry (IHC) — Figure 6. ( A ) Survival outcome analysis in NSCLC patients with wild-type versus mutated PXN. Left and right graphs show overall and individual mutant survival curves, respectively. The relative expression of PXN, p-PXN, BCL-2, FAK, phospho-FAK, Cox IV, DRP-1, and MFN-2 was evaluated by IHC on two sets of samples; seven NSCLC patient samples bearing wild-type PXN and six NSCLC patient samples that were identified to have various PXN mutations as shown in ( B ). ( C and D ) represent the averaged staining intensity scores among all WT and all mutants. ( E ) Shows graph of staining intensity scores of the proteins assayed as expressed in the cytoplasm of the cells. ( F )The panel shows photographs of IHC stains represented in one wild-type and one mutant sample.

Paxillin Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein in Paxillin KO cell line compared to control cell line using Anti-Paxillin Monoclonal Antibody (5H11) (Product # AHO0492). {KO}

Product Details

AHO0492

Applications	Tested Dilution	Publications
Western Blot (WB)	Assay-Dependent	View 6 publications 6 publications
Immunohistochemistry (IHC)	-	View 3 publications 3 publications
Immunohistochemistry (Paraffin) (IHC (P))	1-2 µg/mL	-
Immunocytochemistry (ICC/IF)	2 µg/mL	View 12 publications 12 publications
Immunoprecipitation (IP)	Assay-Dependent	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Dog, Human, Mouse, Rat
Host/Isotype	Mouse / IgG1, kappa
Class	Monoclonal
Type	Antibody
Clone	5H11
Immunogen	Purified recombinant full-length human paxillin protein. View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.2 mg/mL
Purification	Protein G
Storage buffer	PBS, pH 7.4, with 0.2% BSA
Contains	0.09% sodium azide
Storage conditions	4° C
RRID	AB_2536312

Product Specific Information

Staining of formalin-fixed, paraffin-embedded tissues requires boiling tissues in 1mM EDTA, pH 8.0, for 10-20 min followed by cooling at room temperature for 20 min. Recommended positive controls include HeLa cells and colon carcinoma.

Target Information

Paxillin is a focal adhesion protein and a substrate for several tyrosine kinases such as src, FAK, and p210BRC/ABL. The tyrosine phosphorylation of paxillin is affected by conditions that change cell-cell adhesion. Paxillin associates tightly with FAK and Crk through its SH2 domain. This interaction is independent of the extracellular matrix. Although paxillin was initially discovered in fibroblasts, its phosphorylation may also be important during neurite extension during differentiation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

The Lipid-Binding Defective Dynamin 2 Mutant in Charcot-Marie-Tooth Disease Impairs Proper Actin Bundling and Actin Organization in Glomerular Podocytes.

Frontiers in cell and developmental biology

Hamasaki E,Wakita N,Yasuoka H,Nagaoka H,Morita M,Takashima E,Uchihashi T,Takeda T,Abe T,Lee JW,Iimura T,Saleem MA,Ogo N,Asai A,Narita A,Takei K,Yamada H

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunocytochemistry

Sat Jul 16 00:00:00 EDT 2022

The Arf activator GBF1 localizes to plasma membrane sites involved in cell adhesion and motility.

Cellular logistics

Busby T,Meissner JM,Styers ML,Bhatt J,Kaushik A,Hjelmeland AB,Sztul E

AHO0492 was used in Immunocytochemistry-immunoflourescence to investigate the localisation of GBF1 in cell lines derived from human glioblastomas.

Thu Feb 10 00:00:00 EST 2022

Plexin-B2 orchestrates collective stem cell dynamics via actomyosin contractility, cytoskeletal tension and adhesion.

Nature communications

Junqueira Alves C,Dariolli R,Haydak J,Kang S,Hannah T,Wiener RJ,DeFronzo S,Tejero R,Gusella GL,Ramakrishnan A,Alves Dias R,Wojcinski A,Kesari S,Shen L,Sobie EA,Rodrigues Furtado de Mendonça JP,Azeloglu EU,Zou H,Friedel RH

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunocytochemistry

Thu Oct 14 00:00:00 EDT 2021

Plexin-B2 orchestrates collective stem cell dynamics via actomyosin contractility, cytoskeletal tension and adhesion.

Nature communications

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunocytochemistry

Thu Oct 14 00:00:00 EDT 2021

mTORC1 activity is supported by spatial association with focal adhesions.

The Journal of cell biology

Rabanal-Ruiz Y,Byron A,Wirth A,Madsen R,Sedlackova L,Hewitt G,Nelson G,Stingele J,Wills JC,Zhang T,Zeug A,Fässler R,Vanhaesebroeck B,Maddocks ODK,Ponimaskin E,Carroll B,Korolchuk VI

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunocytochemistry

Mon May 03 00:00:00 EDT 2021

Inhibiting FAK-Paxillin Interaction Reduces Migration and Invadopodia-Mediated Matrix Degradation in Metastatic Melanoma Cells.

Cancers

Mousson A,Legrand M,Steffan T,Vauchelles R,Carl P,Gies JP,Lehmann M,Zuber G,De Mey J,Dujardin D,Sick E,Rondé P

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunocytochemistry

Wed Apr 14 00:00:00 EDT 2021

Inhibiting FAK-Paxillin Interaction Reduces Migration and Invadopodia-Mediated Matrix Degradation in Metastatic Melanoma Cells.

Cancers

Mousson A,Legrand M,Steffan T,Vauchelles R,Carl P,Gies JP,Lehmann M,Zuber G,De Mey J,Dujardin D,Sick E,Rondé P

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunocytochemistry

Wed Apr 14 00:00:00 EDT 2021

Fibronectin Patches as Anchoring Points for Force Sensing and Transmission in Human Induced Pluripotent Stem Cell-Derived Pericytes.

Stem cell reports

Iendaltseva O,Orlova VV,Mummery CL,Danen EHJ,Schmidt T

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunocytochemistry

Tue Jun 09 00:00:00 EDT 2020

Nup93 regulates breast tumor growth by modulating cell proliferation and actin cytoskeleton remodeling.

Life science alliance

Bersini S,Lytle NK,Schulte R,Huang L,Wahl GM,Hetzer MW

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunocytochemistry

Wed Jan 01 00:00:00 EST 2020

Vinexin family (SORBS) proteins play different roles in stiffness-sensing and contractile force generation.

Journal of cell science

Ichikawa T,Kita M,Matsui TS,Nagasato AI,Araki T,Chiang SH,Sezaki T,Kimura Y,Ueda K,Deguchi S,Saltiel AR,Kioka N

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunofluorescence

Sun Oct 15 00:00:00 EDT 2017

Vinexin family (SORBS) proteins play different roles in stiffness-sensing and contractile force generation.

Journal of cell science

Ichikawa T,Kita M,Matsui TS,Nagasato AI,Araki T,Chiang SH,Sezaki T,Kimura Y,Ueda K,Deguchi S,Saltiel AR,Kioka N

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunofluorescence

Sun Oct 15 00:00:00 EDT 2017

Adenomatous polyposis coli nucleates actin assembly to drive cell migration and microtubule-induced focal adhesion turnover.

The Journal of cell biology

Juanes MA,Bouguenina H,Eskin JA,Jaiswal R,Badache A,Goode BL

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Western Blot

Mon Sep 04 00:00:00 EDT 2017

Nicotinic acid impairs assembly of leading edge in glioma cells.

Oncology reports

Yang X,Mei S,Niu H,Li J

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Western Blot

Tue Aug 01 00:00:00 EDT 2017

High cell-surface density of HER2 deforms cell membranes.

Nature communications

Chung I,Reichelt M,Shao L,Akita RW,Koeppen H,Rangell L,Schaefer G,Mellman I,Sliwkowski MX

AHO0492 was used in immunocytochemistry and western blot describe the effects of HER2 overexpression in breast cancer cells

Wed Sep 07 00:00:00 EDT 2016

αV-integrins are required for mechanotransduction in MDCK epithelial cells.

PloS one

Teräväinen TP,Myllymäki SM,Friedrichs J,Strohmeyer N,Moyano JV,Wu C,Matlin KS,Muller DJ,Manninen A

AHO0492 was used in immunocytochemistry to demonstrate that αV-integrins regulate the maturation of focal adhesions and cell spreading.

Tue Apr 08 00:00:00 EDT 2014

αV-integrins are required for mechanotransduction in MDCK epithelial cells.

PloS one

Teräväinen TP,Myllymäki SM,Friedrichs J,Strohmeyer N,Moyano JV,Wu C,Matlin KS,Muller DJ,Manninen A

AHO0492 was used in immunocytochemistry to demonstrate that αV-integrins regulate the maturation of focal adhesions and cell spreading.

Tue Apr 08 00:00:00 EDT 2014

Paxillin mutations affect focal adhesions and lead to altered mitochondrial dynamics: relevance to lung cancer.

Cancer biology & therapy

Kawada I,Hasina R,Lennon FE,Bindokas VP,Usatyuk P,Tan YH,Krishnaswamy S,Arif Q,Carey G,Hseu RD,Robinson M,Tretiakova M,Brand TM,Iida M,Ferguson MK,Wheeler DL,Husain AN,Natarajan V,Vokes EE,Singleton PA,Salgia R

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunohistochemistry

Mon Jul 01 00:00:00 EDT 2013

Paxillin mutations affect focal adhesions and lead to altered mitochondrial dynamics: relevance to lung cancer.

Cancer biology & therapy

Published figure using Paxillin monoclonal antibody (Product # AHO0492) in Immunohistochemistry

Mon Jul 01 00:00:00 EDT 2013

c-Yes regulates cell adhesion at the blood-testis barrier and the apical ectoplasmic specialization in the seminiferous epithelium of rat testes.

The international journal of biochemistry & cell biology

Xiao X,Mruk DD,Lee WM,Cheng CY

AHO0492 was used in western blot to elucidate how c-Yes regulates the blood-testis barrier and apical ectoplasmic specialization integrity.

Fri Apr 01 00:00:00 EDT 2011

Presenilin 1 affects focal adhesion site formation and cell force generation via c-Src transcriptional and posttranslational regulation.

The Journal of biological chemistry

Waschbüsch D,Born S,Niediek V,Kirchgessner N,Tamboli IY,Walter J,Merkel R,Hoffmann B

AHO0492 was used in western blot to assess the function of presenilins in the cell-matrix interactions of mouse embryonic fibroblasts

Fri Apr 10 00:00:00 EDT 2009

Loss of caveolin-1 polarity impedes endothelial cell polarization and directional movement.

The Journal of biological chemistry

Beardsley A,Fang K,Mertz H,Castranova V,Friend S,Liu J

Fri Feb 04 00:00:00 EST 2005

Activation of platelet-activating factor receptor-coupled G alpha q leads to stimulation of Src and focal adhesion kinase via two separate pathways in human umbilical vein endothelial cells.

The Journal of biological chemistry

Deo DD,Bazan NG,Hunt JD

AHO0492 was used in Immunoprecipitation to demonstrate that platelet-activating factor activates pertussis toxin-insensitive G alpha(q) protein upon binding to its seven transmembrane receptor.

Fri Jan 30 00:00:00 EST 2004

Aberrant activation of focal adhesion proteins mediates fibrillar amyloid beta-induced neuronal dystrophy.

The Journal of neuroscience : the official journal of the Society for Neuroscience

Grace EA,Busciglio J

Wed Jan 15 00:00:00 EST 2003

Bioinformatics

Protein Aliases: FLJ16691; hCG_1778014; myocardial ischemic preconditioning associated protein 7; PAX-1; PAXI; Paxillin; paxillin alpha; testicular tissue protein Li 134

Gene Aliases: AW108311; AW123232; Pax; PXN

UniProt ID: (Human) P49023, (Mouse) Q8VI36, (Rat) Q66H76

Entrez Gene ID: (Human) 5829, (Mouse) 19303, (Rat) 360820

Molecular Function: actin or actin-binding cytoskeletal protein